siRNA / RNAi /miRNA transfection Rat IEC

Get tips on using IncuCyte® Cell Migration Kit to perform Wound healing assay cell type - human MCF-10A

Get tips on using Iscove’s Modified Dulbecco’s Medium to perform Stem cell Differentiation media iPSCs or hESCs differentiation into cerebellar neuroepithelium (NE)

Get tips on using Unrestricted HD-CGH Microarray ISCA v2, 4x44k to perform Microarray Comperative genomic hybridization - Human PBMCs

Get tips on using IncuCyte® Caspase-3/7 Apoptosis Assay Reagent to perform Apoptosis assay cell type - Caspase 3/7

Get tips on using Corning® 500 mL Iscove’s Modification of DMEM to perform 3D Cell Culture Media hiPSC-derived lung organoids

Get tips on using Iscove's Modified Dulbecco's Medium (IMDM) (ATCC® 30-2005™) to perform Mammalian cell culture media KG1

Plasmid isolation is an important technique in molecular biology or any kind of genetic editing. It involves amplifying plasmids overnight by transforming them into competent bacterial cells. The desired colonies of these bacteria can then be grown in shaker cultures, at appropriate shaking speed, oxygen availability and temperature. These liquid cultures can then be ultracentrifuged to pellet the bacteria, which are then used for plasmid isolation. The bacteria are first resuspended in a buffer, then lysed, neutralized, purified in a column, eluted, precipitated with ethanol and then resuspended. During plasmid isolation, it is important to lyse cells quickly because lysing bacteria for too long may lead to irreversible denaturing of the plasmid. Usually, alkaline lysis is used for isolation because it is a mild treatment. It isolates plasmid DNA and other cell components such as proteins by breaking cells apart with an alkaline solution. Precipitation removes the proteins, and the plasmid DNA recovers with alcohol precipitation. Resuspension and lysis buffers should be mixed thoroughly in order to prevent the DNA from breaking into smaller fragments. This is because broken gDNA can reanneal and remain in the solution, without binding to the column.

Get tips on using Gyros IncSupplier Diversity Partner REXXIP HN BUFFER 25 ML PER VI DFS Item to perform Protein isolation Bacteria - Borrelia burgdorferi

As autophagy is a multi-step process which includes not just the formation of autophagosomes, but most importantly, flux through the entire system, including the degradation upon fusion with lysosomes, which makes it quite challenging for detection. There are several methods for detection in mammalian cells, including immunoblotting analysis of LC3 and p62 and detection of autophagosome formation/maturation by fluorescence microscopy, Currently, there is no single “gold standard” for determining the autophagic activity that is applicable in every experimental context, hence it is recommended to go for the combined use of multiple methods to accurately assess the autophagic activity in any given biological setting.

As autophagy is a multi-step process which includes not just the formation of autophagosomes, but most importantly, flux through the entire system, including the degradation upon fusion with lysosomes, which makes it quite challenging for detection. There are several methods for detection in mammalian cells, including immunoblotting analysis of LC3 and p62 and detection of autophagosome formation/maturation by fluorescence microscopy, Currently, there is no single “gold standard” for determining the autophagic activity that is applicable in every experimental context, hence it is recommended to go for the combined use of multiple methods to accurately assess the autophagic activity in any given biological setting.